Toolbox Cell Viability Test SOP
2011
SOP: Cell Viability Test
Trypan blue exclusion test of cell viability
The dye exclusion test is used to determine the number of viable cells present in a cellsuspension. it
is based on the principle that live cells possess intact cell membranes that exclude certain dyes, such
as trypan blue, whereas dead cells do not
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centrifuge an aliquot of cell suspension being tested for viability 5 min at 150xg
and discard supernatant
resuspend the cell pellet in 10µl Trypan blue solution
place this aliquot on the microscope slide and cover with cover slide
observe the cells under the microscope : dead cells are dyed black, vital cells appear white
Materials:
0.4% trypan blue in PBS (store in dark bottle and filter after prolonged storage)
 160mg Trypan blue in 40 ml PBS